Generation of Inner Ear Sensory Neurons Using Blastocyst Complementation in a Neurog1+/- -Deficient Mouse
This research is the first to produce induced pluripotent stem cell-derived inner ear sensory neurons in the Neurog1 +/- heterozygote mouse using blastocyst complementation. Additionally, this approach corrected non-sensory deficits associated with Neurog1 heterozygosity, indicating that complementation is specific to endogenous Neurog1 function. This work validates the use of blastocyst complementation as a tool to create novel insight into the function of developmental genes and highlights blastocyst complementation as a potential platform for generating chimeric inner ear cell types that can be transplanted into damaged inner ears to improve hearing.
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This is a list of supplementary files associated with this preprint. Click to download.
Supplemental Movie 1. An 8 second video showing segmented and three-dimensionally reconstructed Neurog1+/+ and Neurog1+/- inner ears rotating horizontally, to highlight the morphological differences observed between wild type and non-complemented Neurog1+/- inner ears.
Supplemental Fig 1. Representative gel images of genotyping for a litter from the Neurog1 colony. Top gel is the mutant genotyping, and the bottom gel is the wild type genotyping for the same samples. Wild type (lane 1): 198 bp. Heterozygote (lanes 2-4,6-13): 198 bp and ~ 500 bp. Mutant (lane 5): ~ 500 bp. Heterozygote positive controls (lanes 11-13) were always run in triplicate due to occasional variability in their result. All genotyping was run 3 times for each sample to confirm genotype. Supplemental Fig 2. Representative examples of GFP-expressing induced pluripotent stem cell (iPSC) colonies. (A) GFP expression. (B) Phase Contrast. Note that all of the iPSCs are GFP-labeled. Scale Bar: 100 µm. Supplemental Fig. 3. 2D optical section through a wild type cochlea using sTSLIM. Bracket indicates the sensory cells of the organ of Corti. Note the high degree of resolution throughout the image, in particular the cell bodies in the SGN. Scale Bar: 100 µm. oC: organ of Corti; SGN: Spiral ganglion neurons
Posted 28 Dec, 2020
Received 05 Jan, 2021
Received 31 Dec, 2020
On 27 Dec, 2020
On 25 Dec, 2020
Invitations sent on 24 Dec, 2020
On 08 Dec, 2020
On 08 Dec, 2020
On 08 Dec, 2020
On 15 Nov, 2020
Received 10 Nov, 2020
Received 26 Sep, 2020
On 16 Sep, 2020
On 08 Sep, 2020
Invitations sent on 05 Sep, 2020
On 12 Aug, 2020
On 11 Aug, 2020
On 11 Aug, 2020
On 10 Aug, 2020
Generation of Inner Ear Sensory Neurons Using Blastocyst Complementation in a Neurog1+/- -Deficient Mouse
Posted 28 Dec, 2020
Received 05 Jan, 2021
Received 31 Dec, 2020
On 27 Dec, 2020
On 25 Dec, 2020
Invitations sent on 24 Dec, 2020
On 08 Dec, 2020
On 08 Dec, 2020
On 08 Dec, 2020
On 15 Nov, 2020
Received 10 Nov, 2020
Received 26 Sep, 2020
On 16 Sep, 2020
On 08 Sep, 2020
Invitations sent on 05 Sep, 2020
On 12 Aug, 2020
On 11 Aug, 2020
On 11 Aug, 2020
On 10 Aug, 2020
This research is the first to produce induced pluripotent stem cell-derived inner ear sensory neurons in the Neurog1 +/- heterozygote mouse using blastocyst complementation. Additionally, this approach corrected non-sensory deficits associated with Neurog1 heterozygosity, indicating that complementation is specific to endogenous Neurog1 function. This work validates the use of blastocyst complementation as a tool to create novel insight into the function of developmental genes and highlights blastocyst complementation as a potential platform for generating chimeric inner ear cell types that can be transplanted into damaged inner ears to improve hearing.
Figure 1
Figure 2
Figure 3
Figure 4